5'-Atp → HCID 71405048, compound

HRID 1109509Notes

Reaction equation

HRID 1109509 reaction equation
P(O[CH2:14][C@H:15]1O[C@@H:18](N2C3N=CN=C(N)C=3N=C2)[C@H:17](O)[C@@H:16]1O)(OP(OP(O)(O)=O)(O)=O)(=O)O.N[C@@H:33]([CH2:37][CH2:38][C:39](N[C@H](C(NCC(O)=O)=O)CS)=[O:40])[C:34]([OH:36])=[O:35].[Cl-].[Ca+2].[Cl-]>P([O-])([O-])([O-])=O>[OH:40][C:39]([CH:14]=[CH:15][CH:16]=[CH:17][CH2:18][CH2:14][CH2:15][CH2:16][CH2:17][CH2:18][CH2:14][CH2:15][CH2:16][CH2:17][CH3:18])=[CH:38][CH:37]=[CH:33][C:34]([OH:36])=[O:35]

Participants

5

Reactant 3

HCID 59575'-AtpHCID 124886GlutathioneHCID 5284359Calcium Chloride

Product 1

HCID 71405048HCID 71405048, compound

Reagents, catalysts & solvents 1

HCID 1061PhosphateSolvent

Procedure

5-Lipoxygenase activity was determined according to the method described by Ochi et al., J. Biol. Chem.,258, p. 5754, 1983 using a crude enzyme fraction prepared from peritoneal exudated cells of guinea pigs which were induced by intraperitoneal injection of 2% case in solution. A solution of a test drug was mixed with a reaction solution (2mM ATP, 1 mM glutathione, and the enzyme fraction in 50 mM phosphate buffer, PH 7.4) and the mixture was pre-incubated at 37° C. for 5 minutes. Calcium chloride (1 mM) and 20 μM14C-arachidonic acid were added to the mixture, and then the mixture was subjected to a reaction at 37° C. for 10 minutes. After the completion of the reaction, the product was immediately extracted, dissolved in chloroform/methanol (2:1), and developed by thin-layer chromatography. Radioactivity of the 5-hydroxyeicosatetraenoic acid (5-HETE) fraction was measured using a liquid scintillation counter. Inhibitory rate of each drug-treated group against 5-HETE generation was obtained in comparison with the solvent-treated control, and IC50 was calculated according to the probit method. The results obtained are shown in Table 2 below.

Workup

  1. custom5754, 1983 using a crude enzyme fraction prepared from peritoneal exudated cells of guinea pigs which
  2. customthe mixture was subjected to a reaction at 37° C. for 10 minutes
  3. customAfter the completion of the reaction
  4. extractionthe product was immediately extracted
  5. dissolutiondissolved in chloroform/methanol (2:1)

Sources & evidence

ORD record
ord-263eb419bc744d0eb74db850c12ac224
Source dataset
uspto-grants-1994_06
Patent number
US05321041

Reaction notes

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